Mass spectrometry-based glycomics towards GlycoRNA

Glycosylated RNAs (glycoRNA), a newly discovered and rapidly emerging class of glycosylated biomolecules, challenge the long-standing view that glycosylation occurs exclusively on proteins and lipids. Accumulating evidence indicates that glycoRNA are widely distributed across diverse cell types and species, where they participate in essential biological processes such as immune regulation and intercellular communication. In glycoRNA research, mass spectrometry (MS)-based glycomics provides critical information, offering fundamental insights into its biological functions. This review summarizes recent advances in MS-based analytical strategies for glycoRNA research, including sample collection, RNA extraction, glycoRNA enrichment, glycan release and purification, optional derivatization, liquid chromatography separation, MS detection, and data analysis, with the aim of providing a systematic reference for MS-based RNA glycomics and promoting the further application of glycoRNA in biomedical research and biomarker discovery.